Protein redistribution in TCR-directed NF-kB activation

抗原刺激的T细胞分裂和效应功能的获得依赖于信号 由T细胞受体(TCR)引导的转导，以及随后激活的一些 影响基因表达变化的转录因子。核因子-KB家族的转录因子是 在这一进程中具有至关重要的作用。TCR调节的核因子-KB激活途径将被研究 通过分子、生化和高度先进的成像技术相结合。我们的目标是 建立1)蛋白质-蛋白质相互作用，2)亚细胞蛋白质之间的机制关系 组织和3)TCR导向的核因子-KB中胞质信号转导的生化修饰 路径。这一目标将通过执行三个具体目标来实现： 目的：1.确定信号中定位的蛋白质-蛋白质关联的机制意义 TCR诱导的核因子-kB活化的中间产物。我们将检验这样一个假设，即TCR导向的核因子-KB 信号传递涉及到将已定义的蛋白质复合体有组织地组装和分解成离散的 亚细胞结构域，特定的蛋白质复合体起着关键的机制作用。 目的2.在单个细胞中，研究抗原信号之间的机制关系 强度、信号转导复合体的组装和TCR导向的NF-KB的成功激活 信号转导途径。实验将针对以下假设进行评估： 通过TCR的抗原信号被转换成不激活或完全激活的“二元”结果 效应器功能。此外，这些功能结果依赖于信号转导决定 在单细胞级别上进行的事件。 目的3.明确磷酸化和MALT1相互作用结构域在TCR中的作用。 介导BcHO的降解和活化。我们将进行实验来验证这一假设 BcHO的激活和蛋白水解性破坏是机械耦合的，这两个过程都是 受BcHO与MALT1的相互作用和依赖于PKC的BcHO的磷酸化调节。 这些研究将有助于建立可能导致发展的基础研究基础 新型免疫调节药物，通过高度特异性地抑制抗原受体发挥作用 介导的核因子-kB信号通路的激活。

Antigen-stimulated T cell division and acquisition of effector functions are dependent upon signal transduction directed by the T cell receptor (TCR), and the consequent activation of a number of transcription factors that effect changes in gene expression. Transcription factors of the NF-KB family are of central importance in this process. The TCR-regulated NF-KB activation pathway will be investigated through a combination of molecular, biochemical, and highly advanced imaging techniques. The goal is to establish mechanistic relationships between 1) protein-protein interactions, 2) subcellular protein organization and 3) biochemical modification of cytoplasmic signal transducers in the TCR-directed NF-KB pathway. This goal will be accomplished via the execution of three specific aims: Aim1. Todetermine the mechanistic significance of localizedprotein-protein associations of signaling intermediates in TCR-directed activation of NF-KB. We will test the hypothesis that TCR-directed NF-KB signaling involves the orchestrated assembly and disassembly of defined protein complexes into discrete subcellular domains, with specific protein complexes playing a critical mechanistic role. Aim 2. Toinvestigate, in individual cells, the mechanistic relationship between antigen signal strength, assembly of signal transduction complexes, and successful activation of the TCR-directed NF-KB signal transduction pathway. Experiments will be directed towards the evaluation of the hypothesis that antigen signals through the TCR are converted into "binary" outcomes of either no activation or full activation of effector functions. Furthermore, these functional outcomes are dependent on signal transduction decision events made at the single-cell level. Aim 3. To define the roles of phosphorylation and the MALT1 interaction domain in the TCR- mediated degradation and activation ofBcHO. We will perform experiments to test the hypothesis that activation and proteolytic destruction of BcHO are mechanistically coupled, and that both processes are regulated by BcHO interaction with MALT1 and by PKC-dependent phosphorylation of BcHO. These studies will help establish the basic research foundation that could lead to the development of novel immuno-modulatory drugs, which would function via highly specfic inhibition of antigen-receptor mediated activation of the NF-KB signaling cascade.